Overlay histogram showing SH-SY5Y cells stained with ab119685 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab119685, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

All lanes : Anti-OPA1 antibody [1E81D9] (ab119685) at 1 µg/ml

Lane 1 : whole cell lysates from HeLa cells(human)
Lane 2 : whole cell lysates from H4IIE cells(rat)
Lane 3 : whole cell lysates from MEF cells(mouse)

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP goat anti-mouse at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 112 kDa